The study of hybridization in aquatic plants is complicated by rarity of flower production, absence of roots, and asexuality. Elatine is a cosmopolitan genus of aquatic flowering plants with about 25 species worldwide. Historically, there has been little concern regarding hybridization in the genus due to the prevalence of autogamy (i.e. self-pollination), which potentially limits xenogamous pollen transfer among the species. Two morphologically complex species (Elatine hexandra and E. americana) are the only known polyploids in the genus. In previous phylogenetic analyses, both species resolved incongruently in gene trees obtained from nuclear (ITS) versus plastid (matK/trnK and rbcL) regions. Suspecting that the phylogenetic incongruence might be a consequence of past hybridization events, we tested that hypothesis by conducting an additional phylogenetic analysis of Elatine, which incorporated sequences from a low copy nuclear gene (phyC). Elatine hexandra and E. americana were the only Elatine species exhibiting intraspecific polymorphic sites, i.e. heterozygosity, in phyC. Allele specific amplification enabled us to resolve these polymorphisms for inclusion in a phylogenetic analysis along with the monomorphic phyC sequences within species obtained for the remaining Elatine species. The phyC tree confirmed that both polyploids probably are allopolyploids, in a pattern consistent with the placement of the putative parental taxa in previous phylogenetic analyses of ITS, matK/trnK, and rbcL sequence data. The distributions of E. americana and E. hexandra, along with their potential parental species, are consistent with the proposed hybrid origins for the polyploids and provide additional clues on their geographic regions of origin.